CN117241782A - Sterile aqueous choline salt compositions - Google Patents

Sterile aqueous choline salt compositions Download PDF

Info

Publication number
CN117241782A
CN117241782A CN202280030643.3A CN202280030643A CN117241782A CN 117241782 A CN117241782 A CN 117241782A CN 202280030643 A CN202280030643 A CN 202280030643A CN 117241782 A CN117241782 A CN 117241782A
Authority
CN
China
Prior art keywords
composition
choline
choline chloride
pharmaceutical product
gamma irradiation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202280030643.3A
Other languages
Chinese (zh)
Inventor
代顿·瑞尔丹
杰奎琳·祖莫
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Protara Treatment Co
Original Assignee
Protara Treatment Co
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US17/672,549 external-priority patent/US12083081B2/en
Application filed by Protara Treatment Co filed Critical Protara Treatment Co
Publication of CN117241782A publication Critical patent/CN117241782A/en
Pending legal-status Critical Current

Links

Landscapes

  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Sterile aqueous choline salt compositions, formulations, and methods of use are disclosed. Methods of treating choline deficiency, intestine failure related liver disease (IFALD), and fatty liver disease are also disclosed. Methods of synthesizing choline salts are also disclosed.

Description

Sterile aqueous choline salt compositions
Background
Technical Field
The present disclosure relates to sterile aqueous choline salt compositions, formulations thereof, and methods of use, particularly in the treatment of conditions associated with choline deficiency.
Description of related Art
Choline is a necessary nutrient and a common component of the normal diet, and is generally ingested in the form of phosphatidylcholine found in eggs, meats, nuts and vegetables (Buchman, gateway., 2009,137: S119-S128). The structure of choline includes quaternary amines, which act as methyl donors in many metabolic reactions, similar to B-vitamins and folic acid. Choline is required for cell membrane structure (e.g., phospholipids), transport of triglycerides via Very Low Density Lipoprotein (VLDL) synthesis, transport of cholesterol in bile, intracellular information transfer, brain development and function (e.g., acetylcholine). Choline is critical to cell health and survival: hepatocytes die from apoptosis in choline-deprived media; increased DNA damage and apoptosis have been observed in lymphocytes from normal humans relative to choline deficiency, consistent with increased incidence of liver cancer in rodents after prolonged choline depletion (Shin et al, j. Cell. Biochem.,1997;64:196-208; and da Costa et al, am. J. Clin. Nutr.,2006; 84:88-94).
In vivo 95% of choline is in the form of Phosphatidylcholine (PC). PC is the major lipid of the surface monolayer of VLDL particles and of critical secretory membranes (e.g. ER, golgi, cell membrane). Low PC levels inhibit the packaging and secretion of VLDL (Vance, j.e. and Vance, D.E.,1985,Can.J.Biochem.Cell Biol, 263,12,5898-5909). Exogenous choline is required to maintain adequate PC storage. There are a number of synthetic pathways for PC, but only the de novo pathway is sufficient to supplement PC (Boyer, 2013, compr. Physiol., 3:3). The de novo pathway of PC is CDP-choline, which requires plasma-free choline as a root substrate and is ubiquitous in mammalian tissues (Vance, D.E.,2002,Biochemistry of Lipids,Lipoproteins,and Membranes (Vance, d.e. and Vance, j.e., editions), pages 205-232). In addition to the CDP choline pathway for PC synthesis, the liver also has unique phosphatidylethanolamine methyltransferase (PEMT) activity that provides an alternative pathway for PC synthesis (Noga and Vance D.E.,2003,J.Biol.Chem, 278,24,21851-21859). However, PEMT itself relies in part on choline as a betaine (metabolite of choline) (Sunden, s., renduchentala, m., park, e., miklasz, s., & Garrow, t., 1997) arch. Other choline and PC salvage and resynthesis pathways exist, but only recycle, and do not allow for the production of PC by the recapitulation (Boyer, 2013, comp.
Choline deficiency, which results in reduced levels of phosphatidylcholine, adversely affects a variety of liver and gall functions and can lead to steatosis, bile stasis, and/or hepatocyte death. Steatosis or fatty liver is a broad term describing the accumulation of fat in the liver. Cholestasis is a liver disease that occurs when bile flow from the liver is reduced or blocked. When bile flow changes, it may lead to an accumulation of bilirubin. PC contains about 40% of bile organic matter (Schmitz MGJ, renooij W., gastroent, 1990, 99:1292-1296). Insufficient PC in bile reduces vesicle/mixed micelle formation with cholesterol, increasing free bile salts (Barrios and Lichtenberger,2000, gateway., 118:1179-1186). Free bile salts exert detergent activity on cholangiocytes, restricting bile flow (De Vree, et al, proc. Natl. Acad. Sci. USA,1998, 95:282-287). In addition, choline is an important source of intracellular signaling intermediates (alignment, et al, (2005) cell. Physiol. Biochem.,15 (1-4): 59-68). Choline deficiency induces fragmentation of DNA in cultured hepatocytes (Albright, et al, 1996, FASEBJ,10, 510-516). In addition, hepatocytes die via apoptosis in choline deficient media.
Choline deficiency can lead to liver damage in animals and healthy adults. The experimental choline deficient diet results in a rapid onset of liver abnormalities (e.g., increased liver fat as shown by MRI) that are reversed by normal diet (Zeisel, et al, FASEBJ,1991,5:2093-2098; and Fischer, et al, am. J. Clin. Nutr.,2007, 85:1275-1285). These findings are consistent with those of choline deficiency in several animal species leading to liver steatosis and cirrhosis, skeletal muscle and other organ abnormalities (Patek, et al, proc.Soc.exp.biol.Med.,1975,148:370-374; and reviewed in Buchman, nutr.Clin.practice., 2003, 18:353-358).
Intestinal Failure (IF) occurs when intestinal function falls below the minimum required to absorb large amounts of nutrients and/or water and electrolytes, such that intravenous supplementation is required to maintain health and/or growth. Usually due to surgical excision of the intestine (short bowel syndrome) or diseased, non-functional intestine. IF can be divided into three types: (T-1) it is transient, usually post-operative, and fully reversible; (T-2) it is due to severe disease and requires Parenteral Nutrition (PN) for weeks or months; and (T-3) it requires a long term PN to survive. The underlying cause of T-3 stems from diseases such as cancer, crohn's disease, vascular disease, AIDS, radiation enteritis, etc. (Bakker, H.et al., clin. Nutr.,1999, 18:135-140). Furthermore, a substantial proportion of PN-dependent IF patients develop progressive liver disease or intestinal failure-associated liver disease (IFALD) after survival from Gastrointestinal (GI) disease.
Liver disease in PN-dependent adults and children has been widely observed for decades. Now called IFALD, which is a complication in IF patients, has the greatest risk of mortality (Pironi et al, clin.nutr.,2012, 31:831-45). 47% -65% of adult PN patients suffer from chronic cholestasis (Cavicchi, et al, ann. International. Med.,2000,132:525-532; and Salvino et al, JPEN,2006,30:3, 202-208). 42% of all adult PN patients develop complex liver disease (i.e., extensive portal fibrosis or cirrhosis, bilirubin 3.5mg/dl persists for 1 month, ascites, portal HTN, hepatic encephalopathy or factor V < 50%), and 22% of deaths in PN patients are caused by liver disease (Cavicchi, et al, ann. Intern. Med.,2000,132: 525-532). The pathogenesis is presumed to be multifactorial: PN toxicity (lipids), infections, nutritional deficiencies, including choline deficiency, are all of concern in the literature.
Choline deficiency in PN patients is common and associated with liver injury (Buchman, et al, clin. Nutr.,1993,12:33-37; buchman, gastroent.,2009,137:S119-128; chawlea, et al, gastroent.,1989,97:1514-20; and Burt et al, lancet,1980, month 9, 20, 638-9). Only trace amounts of choline (as an emulsifier) in PN products, impaired from the head synthesis and secondary pathways, choline shortage leading to impaired VLDL fat transport and pathological fat accumulation in hPatoxin, which leads to toxic bile salt accumulation (Noga, et al, J.biol. Chem.,2003,278:21851-21859;Lombardi,et al, J.liquid Res.,1968,9:437-446; yao, et al, J.biol. Chem.,1988,263:2998-3004; and De Vree, et al, proc. Natl. Acad. Sci. USA, 287, 95:282-). Choline is not included in sufficient amounts in PN products that have been approved by the United states society for parenteral and enteral nutrition (ASPEN) as being desirable, but not available as a commercial PN product (Vanek et al, nutr. Clin. Practice., 2012,27 (4), 440-491).
Intravenous (IV) choline administration by IFALD has shown promise in reducing steatosis (Buchman, et al, hepatol, 1995, 22:1399-1403). IV choline administration in patients aged >16 years requiring >80% pn has shown reversal of steatosis and improvement of cholestasis (Buchman, et al, j.part.ent.nutr., 2001, 25:260-268). In addition, IV choline administration is well tolerated in patients (Buchman, et al, clin.Pharm. Ther.,1994, 55:277-83). Parenteral administration of a nutritional solution containing choline salts to a patient has been described as a method of inhibiting fatty liver disease in a human patient (U.S. patent No. 5,567,736).
The U.S. Food and Drug Administration (FDA) requires that all injectable drugs be sterilized by terminal sterilization or by aseptic processing methods. Terminal sterilization is the preferred sterilization method for injectable drugs and is performed after the drug is placed in its primary packaging. Terminal sterilization of aqueous pharmaceuticals may involve heat or irradiation. Selection of the appropriate sterilization method requires a thorough understanding of the physicochemical properties of the drug substance and the properties of the final formulation.
The gamma irradiation process uses cobalt 60, and during radioactive decay cobalt 60 emits gamma rays measured in kilograys (kGy). The energetic gamma radiation interacts with the species by ejecting electrons to form ion pairs, which lead to the formation and excitation of free radicals. Free radicals are highly reactive and can participate in several types of reactions including gas release, double bond formation and cleavage, exchange reactions, electron transfer or crosslinking. In microorganisms, radiation-induced damage may lead to biological changes, which lead to cell death. Although breaking covalent bonds of bacterial DNA is considered a major pathway for cell damage, membrane damage may also contribute significantly in germ cell death. In solution, the molecules may receive energy directly from incident radiation ("direct effect") or in aqueous solutions such as parenteral solutions, by transferring energy from the radiolytic products of water (e.g., hydrogen and hydroxyl radicals and hydrated electrons) to solute molecules ("indirect effect").
It is necessary to examine each new compound to evaluate its radiation stability. Furthermore, for formulated drugs, the stability of the individual components may change when irradiated as part of the product (Jacobs g.p., pharmaceutical Technology,2007 supplement material, phase 2). Due to the "indirect effect," aqueous parenteral drug products present additional challenges in sterilization via gamma irradiation than those in solid form. For example, hydroxyl radicals are the strongest oxidizing species known and are the primary cause of radiation-induced damage to solutes in aqueous irradiation solutions (Sharma et al Advances in Pharmaceutical Product Development and Research,2020, chapter 21, 789-848).
Thus, there is a need for sterile aqueous choline salt compositions and methods of making the same for treating conditions associated with choline deficiency.
Disclosure of Invention
The present disclosure provides sterile choline salt compositions in aqueous media, formulations thereof, and methods of use. The present disclosure also provides methods of treating choline deficiency, intestine failure related liver disease (IFALD), and fatty liver disease.
Drawings
FIG. 1 shows the steam survival curve (R) of spores of Geobacillus stearothermophilus (G.stearothermophilus) suspended in a 50% choline chloride composition 2 0.9895, -1/line slope = D 121 Value = 34 minutes).
Figure 2 shows the serial dilution scheme employed in example 5.
Fig. 3 depicts pH results of "as received" samples of pH studies of 50% w/v choline chloride solution in water for injection (WFI) before and after terminal heat sterilization.
FIG. 4 depicts the pH results of "1:5 diluted" samples of pH studies of 50% w/v choline chloride solution in WFI before and after terminal heat sterilization.
Detailed Description
In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments. However, it will be understood by those skilled in the art that the present invention may be practiced without these details. In other instances, well-known structures have not been shown or described in detail to avoid unnecessarily obscuring the description of the embodiments. Throughout the specification and the claims which follow, unless the context requires otherwise, the word "comprise" and variations such as "comprises" and "comprising" will be interpreted in an open, inclusive sense, i.e. as "including but not limited to". Furthermore, the headings provided herein are for convenience only and do not interpret the scope or meaning of the claimed invention.
Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments. Furthermore, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise. It should also be noted that the term "or" is generally employed in its sense including "and/or" unless the content clearly dictates otherwise. The term "about (about)" should be interpreted to mean plus or minus 10%. For example, "about 5" means 5±0.5. Terms not specifically defined herein should be given their meanings as would be given to those skilled in the art in light of this disclosure and the context.
The following terms used herein have the following meanings, unless otherwise indicated:
"sterile" is defined as free of living microorganisms. "sterilization" is defined as a validated process for rendering a product free of viable microorganisms. The presence of microorganisms is expressed as a probability. Although the probability can be reduced to a very low number, it can never be reduced to zero with certainty. Thus, the term "Sterility Assurance Level (SAL)" is used as a measure of sterility. "Sterility Assurance Level (SAL)" refers to the probability of the presence of viable microorganisms on a product after sterilization and is generally expressed as 10 -n . SALs can also be used to describe populations of microorganisms that are destroyed by the sterilization process, although this is different from the definition of probability. Commonly referred to as "log reduction" (a technical reduction of one order of magnitude), represents a 90% reduction in the microbial population. Thus, a "6-log reduction" is achieved (10 -6 ) In theory, will reduce the initial population of 100 tens of thousands of organisms to very close to zero. Sterility may also be expressed by the presence of "Colony Forming Units (CFU)" which is used to describe the visible growth of microorganisms produced from a single cell or multiple cells.
"choline salt" refers to a class of quaternary ammonium salts containing an N, N, N-trimethylethanolamine cation and the corresponding counter anion, which can be represented by the general formula wherein X - Representing the corresponding counter anion:
suitable counter anions include, but are not limited to, halides, such as chloride Cl - And hydrogen tartrate ((2 r,3 r) -2,3, 4-trihydroxy-4-oxobutyrate). The choline salts can be prepared from inorganic acids or from organic acids. Examples of the inorganic acid include hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid, and phosphoric acid. Suitable organic acids may be selected from the aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic acid, acetic acid, propionic acid, succinic acid, ethanolAcid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, anthranilic acid, 4-hydroxybenzoic acid, phenylacetic acid, mandelic acid, hippuric acid, malonic acid, oxalic acid, pamoic acid (pamoic), methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, pantothenic acid, trifluoromethanesulfonic acid, 2-hydroxyethanesulfonic acid, p-toluenesulfonic acid, sulfanilic acid, cyclohexylsulfamic acid, stearic acid, alginic acid, β -hydroxybutyric acid, salicylic acid, galactaric acid, and galacturonic acid.
An "aqueous medium" is defined as a liquid that is greater than about 50% water.
"weight/volume%" is a means of indicating the concentration of a solution, wherein
"Water for injection (WFI)" is a sterile, pyrogen-free aqueous formulation for injection that does not contain bacteriostats, antimicrobials or added buffers.
"bacteriostatic agent" refers to a substance that prevents the proliferation of bacteria without destroying the bacteria.
"antimicrobial agent" refers to a natural or synthetic substance that kills or inhibits the growth of microorganisms such as bacteria, fungi, and algae.
"preservative" refers to an antimicrobial component added to the composition to help maintain the safety of the composition by inhibiting the growth of microbial contaminants or reducing the number of microbial contaminants, or both.
"amino acid" is a compound containing a carboxyl group (-COOH) and an amino group (-NH) 2 ) Simple organic compounds of groups. The term "amino acid" may refer to naturally occurring or synthetic (i.e., man-made) amino acids.
"vitamins" refers to any one of a group of organic compounds that are critical to normal growth and nutrition and are required in small amounts in the diet because they cannot be synthesized by the body. Examples of vitamins may include, but are not limited to, vitamin a (as all-trans retinol, all-trans retinyl esters, and all-trans beta-carotene and other provitamin a carotenoids), vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B6 (pyridoxine), vitamin B7 (biotin), vitamin B9 (folic acid or folate), vitamin B12 (cobalamin), vitamin C (ascorbic acid), vitamin D (calciferol), vitamin E (tocopherols and tocotrienols), and vitamin K (quinones).
"fatty acid" refers to any of the carboxylic acids consisting of hydrocarbon chains and terminal carboxyl groups, especially those present as esters in fats and oils. Fatty acids may include, but are not limited to, alpha-linolenic acid, linoleic acid, docosahexaenoic acid, and gamma-linolenic acid.
"ionic strength" is the total ion concentration in a solution. The ionic strength is calculated by the formula Calculating, wherein c i Is the molar concentration (mol/L) of ion I, z i Is the charge number of the ion and the sum is taken to be the sum of all ions in the solution. For example, for a 1:1 electrolyte such as choline chloride, where each ion is singly charged, the ionic strength is equal to the sum of the concentrations of each ion (choline +1 Chloride compound 1 )。
By "substantially free of microorganisms, bacteria, or specific strains of bacteria, such as staphylococcus aureus (s. Aureus) or geobacillus stearothermophilus (Geobacillus stearothermophilus)" is meant bacteria having a low (i.e.,.ltoreq.1 colony forming unit, CFU/mL) but clinically acceptable level. Microorganisms include viruses, bacteria, archaea, fungi, plants such as algae and protozoa.
By "antimicrobial growth" is meant that the composition meets the criteria set forth, for example, in the U.S. food and drug administration and U.S. pharmacopoeia, for products prepared using aqueous bases or vehicles. For example, for bacteria, antimicrobial growth may mean that the initial calculated count is not less than 1.0log reduced at 7 days, not less than 3.0log reduced at 14 days, and not increased at 28 days as compared to the 14 day count. For example, for yeasts and molds, antimicrobial growth may mean that at 7, 14 and 28 days there is no increase compared to the initially calculated count.
The "D value" or decimal reduction time (or decimal reduction dose) is the time (or dose) required to achieve a logarithmic reduction, i.e., kill 90% (or 1log or more) of the relevant microorganisms under a given condition (e.g., temperature) or set of conditions.
"chemical stability" refers to the resistance of a substance to degradation into its known or unknown degradation products. For example, trimethylamine is a known degradation product of choline chloride. Acceptable trimethylamine levels can be as high as 0.2%.
By "pharmaceutical product" is meant a finished dosage form, such as a tablet, capsule, solution, etc., containing an active pharmaceutical ingredient, typically, but not necessarily, in combination with a non-active ingredient.
In certain embodiments of the present invention, a pH adjuster may be added to the composition. The choice of pH modifier may affect resistance to microbial excitation and/or stability of choline salts, as measured by the reduction of measurable degradation products. The pH adjuster may include an acid such as malic acid, citric acid, acetic acid, boric acid, lactic acid, hydrochloric acid, phosphoric acid, sulfuric acid, sulfonic acid, or nitric acid. The pH adjuster may also include a base such as acetanilide, ammonia, calcium hydroxide, potassium bicarbonate, potassium hydroxide, sodium bicarbonate, sodium dihydrogen phosphate, sodium citrate, sodium tartrate (sodium tartrate), sodium carbonate, sodium hydroxide, thiourea, or urea. Any pH adjuster disclosed herein or known to one of ordinary skill in the art is contemplated herein.
As used herein, a "pharmaceutical composition" is synonymous with a composition.
By "pharmaceutically acceptable" is meant molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions upon administration to a subject. As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. To the extent that any conventional medium or agent is incompatible with the active ingredient, its use in a therapeutic composition is unsuitable. Additional compatible active ingredients may be incorporated into the composition. For human administration, the formulation should meet sterility, pyrogenicity, general safety, and purity standards as required by the U.S. Food and Drug Administration (FDA). Conventional procedures and ingredients for selecting and preparing suitable compositions are described, for example, in Remington: the Science and Practice of Pharmacy, 21 st edition, gennaro, eds., lippencott Williams & Wilkins (2005) and The United States Pharmacopeia: the National Formulary (USP 36NF 31) published 2013.
"excipient" refers to certain embodiments that are added as a diluent (where "diluent" refers to a substance used to dilute something) or vehicle, or to provide form or consistency that is more or less an inert substance. Excipients may also enhance resistance to microbial growth and thus act as preservatives. Such excipients include, but are not limited to, xylitol, mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose derivatives, magnesium carbonate, and the like.
As used herein, the term "effective amount" refers to an amount of a particular agent sufficient to achieve a desired effect in a subject receiving treatment with the agent. Desirably, an effective amount of the agent is an amount sufficient to inhibit or treat the disease without substantial toxicity to the subject. The effective amount of the agent will depend on the subject being treated, the severity of the affliction, and the manner of administration of the pharmaceutical composition. One of ordinary skill in the art, in light of the present disclosure, will understand the method of determining an effective amount of the disclosed agent sufficient to achieve a desired effect in a subject.
As used herein, the term "chronic" refers to a medical disorder or condition that persists or frequently recurs over time.
The dosage form may be administered once a day, or more than once a day, such as two or three times a day. Alternatively, if prescription information for the prescribing physician or drug is found to be appropriate, the dosage form may be administered less frequently than daily, such as every other day or weekly. The dosing regimen includes, for example, dose titration to the extent necessary or useful for the indication to be treated, thereby allowing the patient's body to adapt to the treatment, to minimize or avoid undesired treatment-related side effects, and/or to maximize the therapeutic effect of the treatment. Suitable dosage regimens and/or forms include, for example, those set forth in the latest version of the Physics' desk reference, which is incorporated herein by reference.
"direct injection" refers to intravenous injection, wherein the substance is injected directly into the vein.
"indirect injection" refers to intravenous injection in which a substance is introduced into another source prior to direct injection into a vein. For example, without limiting the possible routes of indirect injection, indirect injection includes introducing a substance into an IV bag, where the contents of the IV bag are then administered to a subject via injection into a vein as the end of travel (e.g., the contents of an IV bag, with or without the substance to be introduced, may flow through or be contained in other channels or devices prior to final injection into the vein).
As used herein, "kit" refers to any combination of a number of items. For example, a kit may include a composition and a syringe for injecting the composition. The kit may also include a preloaded syringe for injection.
"gamma irradiation" refers to gamma irradiation of a material, wherein high energy photons are emitted from an isotope source (e.g., cobalt 60). High energy gamma radiation generates electron destruction (ionization) in any material it encounters. In living cells, these disruptions can lead to damage to DNA and other cellular structures. Changes in these photons caused at the molecular level may lead to death or failure of the organism. This effect may be used to kill bacteria, insects or other living contaminants that may be present in or on the material.
"Choline deficiency" refers to clinically defined deficiency of choline. Choline is a nutrient essential to humans and is essential for the normal function of all cells. As a key component of the cell membrane, it ensures the structural integrity and signaling function of the cell. Choline is also used for neurotransmission (acetylcholine as a metabolite), is a major source of methyl donors and is essential for lipid transport from the liver. Given these many different roles, choline deficiency can lead to disorders in many bodily systems, including liver, muscle and lymphocytes in humans, and in addition kidneys, pancreas and developing brain and nervous system in animals. Choline deficiency may be characterized by <7nmol of free choline.
By "Parenteral Nutrition (PN)" is meant the intravenous administration of nutrition, which may include proteins, carbohydrates, fats, minerals and electrolytes, vitamins and other trace elements, to maintain a good nutritional status, to patients who are unable to eat or absorb sufficient food either by gavage or by the oral cavity.
"parenteral support" includes administration of parenteral fluids alone or in combination with parenteral nutritional solutions.
By "Intestinal Failure (IF)" is meant that the channel function falls below the minimum level required to absorb large amounts of nutrients and/or water and electrolytes, such that intravenous supplementation is required to maintain health and/or growth. In some cases, IF may be due to surgical excision of the intestine (short bowel syndrome) or diseased, non-functional intestine.
"intestinal failure-related liver disease (IFALD)" refers to liver disease associated with intestinal failure, which may occur in PN-dependent subjects. In some cases, IFALD develops in chronic intestinal failure patients who receive parenteral nutrition for a long period of time and may be characterized by liver steatosis and/or cholestasis, and may be accompanied by one or more other signs of liver injury, including but not limited to elevated liver function tests, fibrosis, cirrhosis, and end-stage liver disease (ESLD). In some cases, IFALD may have been previously referred to as PN-associated liver disease (PNALD).
"fatty liver disease" refers to a condition characterized by excessive accumulation of lipids (fat) in the liver. The accumulation of fat in the liver results in a series of clinical manifestations and progresses in stages. Depending on the etiology, each stage may be characterized as non-alcoholic or alcoholic. Progress begins with simple fatty liver or steatosis. This stage is generally considered benign and is characterized by an increased appearance of fat in the liver. Fatty liver may be characterized as non-alcoholic (NAFL) or Alcoholic (AFL). The next stage of fatty liver disease is a form of hepatitis known as steatohepatitis, which is characterized by further accumulation of fat and inflammation of liver tissue. Steatohepatitis can be Nonalcoholic (NASH) or Alcoholic (ASH). NASH and ASH may both lead to the next stage of fatty liver disease, NASH-related or ASH-related fibrosis, respectively, characterized by liver scarring. Finally, fibrosis may progress to cirrhosis, which causes irreversible damage to the liver, and is the most severe stage. Cirrhosis may be nonalcoholic or alcoholic.
"type 1 glass" refers to borosilicate glass having good chemical resistance. Type 1 glass is used for pharmaceutical products requiring a minimum of reaction vessels. Exemplary products include, but are not limited to, glass vials, filled syringes, cartridges, and ampoules.
"Delamination Control (DC) glass" refers to glass that is resistant to delamination (i.e., degradation of the surface glass). Exemplary products include, but are not limited to, glass vials, filled syringes, cartridges, and ampoules.
"pure ethanol" refers to ethanol of 200 gauge (100%).
When referring to 2-chloroethanol or aluminum, "substantially free of (non) detectable" means that the amount of either material is within 10% of the limit of detection.
In some embodiments, the invention provides a sterile composition comprising a choline salt in an aqueous medium.
In certain embodiments, the composition contains 25% to 75% choline salt by weight/volume%. In a specific embodiment, the composition contains 50% choline salt by weight/volume%.
In some embodiments, the choline salt is choline chloride. In other embodiments, the choline salt is choline bitartrate.
In some embodiments, the present invention provides a sterile composition for intravenous injection comprising choline chloride in an aqueous medium, wherein the choline chloride is present in the composition at a level of 25% to 75% choline chloride salt by weight/volume%.
In some embodiments, the invention provides a sterile composition for intravenous injection comprising choline chloride in an aqueous medium, wherein the choline chloride is present in the composition at a level of 50% choline chloride salt by weight/volume%.
In some embodiments, the present invention provides a sterile composition for intravenous injection due to the composition of choline chloride in an aqueous medium, wherein the choline chloride is present in the composition at a level of 50% choline chloride by weight/volume%.
In some embodiments, the aqueous medium is water for injection.
In certain embodiments, the composition does not contain a preservative. In other embodiments, the composition contains a preservative. In some embodiments, the composition contains at least one amino acid, at least one vitamin, and/or at least one fatty acid. In some embodiments, the composition contains at least one amino acid. In other embodiments, the composition contains at least one vitamin. In certain embodiments, the composition contains at least one fatty acid. In other embodiments, the composition contains a pharmaceutically acceptable carrier, diluent or excipient.
In some embodiments, the composition has an ionic strength of at least 0.3 moles per liter (molar). In certain embodiments, the composition has an ionic strength of 0.3 to 7 moles per liter (molar). In certain specific embodiments, the composition has an ionic strength of about 7 moles per liter (molar).
In some embodiments, the composition has a pH of about 4 to 7.
In some embodiments, the composition is substantially free of microorganisms. In some embodiments, the composition is substantially free of bacteria.
In some embodiments, the composition is substantially free of staphylococcus aureus, geobacillus stearothermophilus, andand/or Bacillus pumilus (B.pumilus). In certain embodiments, the composition contains less than 10 -1 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus. In certain embodiments, the composition contains less than 10 - 2 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus. In certain embodiments, the composition contains less than 10 -3 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus. In certain embodiments, the composition contains less than 10 -4 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus. In certain embodiments, the composition contains less than 10 -5 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus. In certain embodiments, the composition contains less than 10 -6 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus.
In some embodiments, the composition is substantially free of staphylococcus aureus. In certain embodiments, the composition contains less than 10 -1 CFU/mL staphylococcus aureus. In certain embodiments, the composition contains less than 10 -2 CFU/mL staphylococcus aureus. In certain embodiments, the composition contains less than 10 -3 CFU/mL staphylococcus aureus. In certain embodiments, the composition contains less than 10 -4 CFU/mL staphylococcus aureus. In certain embodiments, the composition contains less than 10 -5 CFU/mL staphylococcus aureus. In certain embodiments, the composition contains less than 10 -6 CFU/mL staphylococcus aureus.
In some embodiments, the composition is substantially free of geobacillus stearothermophilus. In certain embodiments, the composition contains less than 10 -1 CFU/mL Geobacillus stearothermophilus. In certain embodiments, the composition contains less than 10 -2 CFU/mL Geobacillus stearothermophilus. In certain embodiments, the composition contains less than 10 -3 CFU/mL thermophilic fat plotBacillus. In certain embodiments, the composition contains less than 10 -4 CFU/mL Geobacillus stearothermophilus. In certain embodiments, the composition contains less than 10 -5 CFU/mL Geobacillus stearothermophilus. In certain embodiments, the composition contains less than 10 -6 CFU/mL Geobacillus stearothermophilus.
In some embodiments, the composition is substantially free of bacillus pumilus. In certain embodiments, the composition contains less than 10 -1 CFU/mL Bacillus pumilus. In certain embodiments, the composition contains less than 10 -2 CFU/mL Bacillus pumilus. In certain embodiments, the composition contains less than 10 -3 CFU/mL Bacillus pumilus. In certain embodiments, the composition contains less than 10 -4 CFU/mL Bacillus pumilus. In certain embodiments, the composition contains less than 10 -5 CFU/mL Bacillus pumilus. In certain embodiments, the composition contains less than 10 -6 CFU/mL Bacillus pumilus.
In some embodiments, the composition has at least 10 -3 -10 -6 Sterility assurance level of (c). In certain embodiments, the composition has a weight ratio of at least 10 -3 Sterility assurance level of (c). In other embodiments, the composition has a weight ratio of at least 10 -4 Sterility assurance level of (c). In other embodiments, the composition has a weight ratio of at least 10 -5 Sterility assurance level of (c). In other embodiments, the composition has a weight ratio of at least 10 -6 Sterility assurance level of (c).
In some embodiments, the composition is sterilized by the ionic strength of the composition. In some embodiments, the composition is sterilized by gamma irradiation. In other embodiments, the composition is sterilized by a combination of ion intensity and gamma irradiation. In some embodiments, the gamma irradiation is at least 20kGy. In some embodiments, the gamma irradiation is 18-25kGy. In some embodiments, the gamma irradiation is 25-33kGy. In some embodiments, the gamma irradiation is 45-59kGy.
In some embodiments, the composition is suitable for administration via indirect injection or via direct injection. In particular embodiments, the compositions are suitable for administration via direct injection. In other specific embodiments, the composition is suitable for administration via indirect injection.
In some embodiments, the composition is resistant to microbial growth. In some embodiments, the composition is chemically stable.
In some embodiments, the present invention provides methods of preparing a composition comprising combining a choline salt with an aqueous medium and adjusting the concentration of the choline salt in the aqueous medium.
In some embodiments, the pH is also adjusted. In certain embodiments, the pH adjuster is an acid. In particular embodiments, the acid is malic acid, citric acid, acetic acid, boric acid, lactic acid, hydrochloric acid, phosphoric acid, sulfuric acid, sulfonic acid, or nitric acid.
In other embodiments, the pH adjuster is a base. In other embodiments, the base is acetanilide, ammonia, calcium hydroxide, potassium bicarbonate, potassium hydroxide, sodium bicarbonate, sodium dihydrogen phosphate, sodium citrate, sodium tartrate (sodium tartrate), sodium carbonate, sodium hydroxide, thiourea, or urea.
In some embodiments, the method comprises sterilizing the composition by its ionic strength. In some embodiments, the method comprises sterilizing the composition by gamma irradiation. In other embodiments, the method comprises sterilizing the composition by a combination of ion intensity and gamma irradiation.
In some embodiments, the gamma irradiation is at least 20kGy. In some embodiments, the gamma irradiation is 18-25kGy. In some embodiments, the gamma irradiation is 25-33kGy. In some embodiments, the gamma irradiation is 45-59kGy.
In some embodiments, the present invention provides a method of preparing a composition as described herein, comprising combining choline chloride with water for injection to prepare a 50% (w/v) solution having an ionic strength of about 7M and a pH between about 4-7, and exposing the solution to gamma radiation to produce at least 10 -6 Sterility assurance level of (c).
In some embodiments, the gamma irradiation is at least 20kGy. In some embodiments, the gamma irradiation is 18-25kGy. In some embodiments, the gamma irradiation is 25-33kGy. In some embodiments, the gamma irradiation is 45-59kGy.
In some embodiments, the invention provides a composition as described herein, prepared by a method as described herein.
In some embodiments, the present invention provides compositions prepared by combining a choline salt and water for injection such that the ionic strength of the composition facilitates sterilization of the composition, and further sterilizing the composition by exposure to gamma radiation. In a specific embodiment, the composition contains 50% choline chloride by weight/volume%.
In some embodiments, the composition is filtered through a micron filter. In some embodiments, the composition is filtered through two micron filters in series. In some embodiments, the composition is filtered through a 0.2 micron filter. In some embodiments, the composition is filtered through two 0.2 micron filters in series. In some embodiments, the composition is filtered through a 0.45 micron filter. In some embodiments, the composition is filtered through two 0.45 micron filters in series.
In some embodiments, the invention provides a method of treating choline deficiency in a subject comprising administering to the subject an effective amount of a composition as described herein.
In some embodiments, the invention provides methods of providing parenteral support to a subject comprising administering to the subject an effective amount of a composition as described herein.
In some embodiments, the invention provides methods of providing parenteral nutrition to a subject comprising administering to the subject an effective amount of a composition as described herein.
In some embodiments, the invention provides a method of treating liver and gall juice stasis in a subject comprising administering to the subject an effective amount of a composition as described herein.
In some embodiments, the invention provides a method of treating liver steatosis in a subject comprising administering to the subject an effective amount of a composition as described herein.
In some embodiments, the invention provides a method of treating an intestinal failure-related liver disease (IFALD) in a subject, comprising administering to the subject an effective amount of a composition as described herein.
In some embodiments, the invention provides a method of treating fatty liver disease in a subject comprising administering to the subject an effective amount of a composition as described herein. In some embodiments, the fatty liver disease is AFL, ASH, NAFL, NASH, NASH-associated liver fibrosis or ASH-associated liver fibrosis. In some embodiments, the fatty liver disease is Alcoholic Fatty Liver (AFL). In some embodiments, the fatty liver disease is Alcoholic Steatohepatitis (ASH). In some embodiments, the fatty liver disease is non-alcoholic fatty liver disease (NAFL). In some embodiments, the fatty liver disease is nonalcoholic steatohepatitis (NASH). In some embodiments, the fatty liver disease is NASH-related liver fibrosis. In some embodiments, the fatty liver disease is ASH-related liver fibrosis.
In some embodiments, the treatment comprises administering to the subject a composition described herein as parenteral support. Parenteral support includes administration of parenteral fluids alone or in combination with parenteral nutritional solutions. For example, when only a fluid is administered, a solution comprising a composition described herein may be provided by administration as a separate Y-line. In some embodiments, the treatment comprises administering a composition described herein to the subject as part of a parenteral support solution, wherein administration occurs at least once per day or as determined by the treating physician. In some embodiments, the treatment comprises administering a composition described herein to the subject as part of a parenteral support solution, wherein the administration occurs at least once per day or on a schedule basis to obtain normal plasma choline levels as determined by the treating physician. For example, the infusion time may be several hours (e.g., 10-14 hours), which may be administered continuously, or may be discontinued during one or more dosing intervals.
In some embodiments, the treatment comprises administering a composition described herein to a subject as part of a parenteral nutritional solution. In some embodiments, the treatment comprises administering a composition described herein to the subject as part of a parenteral nutritional solution, wherein administration occurs at least once per day or as determined by the treating physician. In some embodiments, the treatment comprises administering a composition described herein to the subject as part of a parenteral nutrition solution, wherein administration occurs at least once per day or on a schedule basis to obtain normal plasma choline levels as determined by the treating physician. For example, the infusion time may be several hours (e.g., 10-14 hours), which may be administered continuously, or may be discontinued during one or more dosing intervals.
In particular embodiments, the compositions as described herein are administered to a subject via direct injection. In other specific embodiments, the compositions as described herein are administered to a subject via indirect injection.
In some embodiments, the present invention provides choline salt compositions, wherein the composition is sterilized in liquid form by gamma irradiation of at least 25kGy and is the irradiation product of a composition comprising 25% -75% choline salt by weight/volume% in an aqueous medium, wherein the composition has: (a) a pH of about 4-7; (b)>0.3M ionic strength; (c) at least 10 -3 -10 -6 Sterility assurance level of (c). In a specific embodiment, the ionic strength of the composition is about 7M. In particular embodiments, the composition has a sterility assurance level of at least 10 -6 M. In a specific embodiment, the aqueous medium is water for injection. In a specific embodiment, the choline salt is choline chloride. In a specific embodiment, the composition comprises 50% choline salt by weight/volume%.
In some embodiments, the composition is used in a method of treating choline deficiency in a subject comprising administering to the subject an effective amount of a composition as described herein. In some embodiments, the composition is used in a method of treating a defect in a subject in parenteral support comprising administering to the subject an effective amount of a composition as described herein. In some embodiments, the composition is used in a method of treating a defect in a subject in parenteral nutrition comprising administering to the subject an effective amount of a composition as described herein. In some embodiments, the composition is used in a method of treating liver steatosis in a subject, comprising administering to the subject an effective amount of a composition as described herein. In some embodiments, the composition is used in a method of treating liver and gall stasis in a subject, comprising administering to the subject an effective amount of a composition as described herein. In some embodiments, the composition is used in a method of treating an intestinal failure-related liver disease (IFALD) in a subject, comprising administering to the subject an effective amount of a composition as described herein. In some embodiments, the composition is used in a method of treating fatty liver disease in a subject comprising administering to the subject an effective amount of a composition as described herein. In some embodiments, the fatty liver disease is AFL, ASH, NAFL, NASH, NASH-associated liver fibrosis or ASH-associated liver fibrosis.
In some embodiments, the present invention provides a composition as described herein, wherein the composition is packaged in the form of a vial for injection.
In some embodiments, the composition is packaged in a glass container. In some embodiments, the composition is packaged in a type 1 glass container. In other embodiments, the composition is packaged in a layered controlled (DC) glass container.
In some embodiments, the present invention provides a composition as described herein, wherein the composition is packaged in the form of an IV bag.
In some embodiments, the present invention provides a composition as described herein, wherein the composition is packaged in the form of a preloaded syringe.
In some embodiments, the invention provides a composition as described herein, wherein the composition is packaged in the form of a kit comprising the composition and a syringe. In a specific embodiment, the kit comprises the composition packaged in a glass container. In a specific embodiment, the kit comprises the composition packaged in a type 1 glass container. In particular embodiments, the kit comprises the composition packaged in a layered-controlled (DC) glass container. In particular embodiments, the kit comprises the composition packaged in an IV bag. In other specific embodiments, the kit comprises the composition packaged as a preloaded syringe.
In some embodiments, the present invention provides compositions and packages as described herein, wherein the compositions and packages are sterilized by gamma irradiation. In some embodiments, the present invention provides compositions and packages as described herein, wherein the compositions are sterilized by ion intensity and gamma irradiation, and the packages are sterilized by gamma irradiation. In a specific embodiment, the composition and package are sterilized by gamma irradiation of at least 20 kGy. In some embodiments, the compositions and packages are sterilized by gamma irradiation of 18-25 kGy. In some embodiments, the composition and package are sterilized by gamma irradiation of 25-33 kGy. In some embodiments, the compositions and packages are sterilized by gamma irradiation of 45-59 kGy.
In some embodiments, the invention provides a sterile, ready-to-use pharmaceutical composition of choline chloride comprising: a primary packaging container containing a sterile aqueous choline chloride solution having a concentration of 25-75% choline chloride in weight/volume% in an aqueous medium; a seal for sealing the primary packaging container; wherein the choline chloride solution is according to at least 10 -3 -10 -6 No viable microbial contamination. In a specific embodiment, the aqueous choline chloride solution has a choline chloride concentration of 50% by weight/volume%. In particular embodiments, the sterility assurance level is at least 10 -6 . In certain embodiments, the aqueous medium is water for injection. In some embodiments, the primary package is a glass container. In some embodiments, the primary package is a type 1 glass container. In other embodiments, the primary package is a DC glass container. In other embodimentsIn this case, the primary is packaged in an IV bag. In other embodiments, the primary package is a preloaded syringe.
In some embodiments, the ready-to-use pharmaceutical composition is sterilized by gamma irradiation. In a specific embodiment, the ready-to-use pharmaceutical composition is sterilized by gamma irradiation of at least 20 kGy. In other specific embodiments, the ready-to-use pharmaceutical composition is sterilized by gamma irradiation of 25-33 kGy. In other specific embodiments, the ready-to-use pharmaceutical composition is sterilized by gamma irradiation of 45-59 kGy.
In some embodiments, the ready-to-use pharmaceutical composition is suitable for administration via indirect injection. In other embodiments, the ready-to-use drug is suitable for administration via direct injection. In other embodiments, the ready-to-use drug may be stable in the primary package for at least 3 months.
In some embodiments, the invention provides a sterile aqueous choline chloride pharmaceutical product prepared by the process of: (i) Dissolving choline chloride in water for injection to a final concentration of 40% -60% choline chloride in weight/volume%; (ii) filtering the solution through a micron filter; (iii) transferring the solution to a glass vial; (iv) sealing the vial; (v) The pharmaceutical product is sterilized using gamma irradiation. In certain embodiments, in step (ii), the sterile aqueous choline chloride pharmaceutical product is filtered through a 0.2 micron filter. In certain embodiments, in step (ii), the sterile aqueous choline chloride pharmaceutical product is filtered through a 0.45 micron filter. In certain embodiments, in step (ii), the sterile aqueous choline chloride pharmaceutical product is filtered through two micron filters in series.
In some embodiments, the invention provides a sterile aqueous choline chloride pharmaceutical product prepared by the process of: (i) Dissolving choline chloride in water for injection to a final concentration of 40% -60% choline chloride in weight/volume%; (ii) filtering the solution through a 0.2 micron filter; (iii) transferring the solution to a glass vial; (iv) sealing the vial; (v) The pharmaceutical product is sterilized using gamma irradiation.
In a specific embodiment, the sterile aqueous choline chloride pharmaceutical product contains 50% choline chloride by weight/volume%. In certain embodiments, the sterile aqueous choline chloride pharmaceutical product is filtered through two 0.2 micron filters in series. In certain embodiments, the sterile aqueous choline chloride pharmaceutical product is filtered through two 0.45 micron filters in series. In other embodiments, the sterile aqueous choline chloride pharmaceutical product is sterilized by gamma irradiation of at least 20 kGy. In other specific embodiments, the sterile aqueous choline chloride pharmaceutical product is sterilized by gamma irradiation of 25-33 kGy. In other specific embodiments, the sterile aqueous choline chloride pharmaceutical product is sterilized by gamma irradiation of 45-59 kGy.
In some embodiments, the sterile aqueous choline chloride pharmaceutical product is transferred to a vial, wherein the vial is a glass container. In some embodiments, the sterile aqueous choline chloride pharmaceutical product is transferred to a vial, wherein the vial is a type 1 glass container. In some embodiments, the sterile aqueous choline chloride pharmaceutical product is transferred to a vial, wherein the vial is a DC glass container. In a specific embodiment, the sterile aqueous choline chloride pharmaceutical product is sealed in a vial, wherein the vial is sealed with a rubber stopper. In other specific embodiments, the sterile aqueous choline chloride pharmaceutical product is sealed in a vial, wherein the vial is further sealed with an aluminum gland.
In certain embodiments, the pharmaceutical product does not contain a preservative. In other embodiments, the pharmaceutical product contains a preservative. In some embodiments, the pharmaceutical product contains at least one amino acid, at least one vitamin, and/or at least one fatty acid. In some embodiments, the pharmaceutical product contains at least one amino acid. In other embodiments, the pharmaceutical product contains at least one vitamin. In certain embodiments, the pharmaceutical product contains at least one fatty acid. In other embodiments, the pharmaceutical product contains a pharmaceutically acceptable carrier, diluent or excipient.
In some embodiments, the pharmaceutical product has an ionic strength of at least 0.3 moles per liter (molar). In certain embodiments, the pharmaceutical product has an ionic strength of 0.3 to 7 moles per liter (molar). In certain specific embodiments, the pharmaceutical product has an ionic strength of about 7 moles per liter (molar).
In some embodiments, the pharmaceutical product has a pH of about 4-7.
In some embodiments, the pharmaceutical product is substantially free of microorganisms. In some embodiments, the pharmaceutical product is substantially free of bacteria.
In some embodiments, the pharmaceutical product is substantially free of staphylococcus aureus, geobacillus stearothermophilus, and/or bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -1 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -2 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -3 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -4 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -5 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -6 CFU/mL of Staphylococcus aureus, geobacillus stearothermophilus and/or Bacillus pumilus.
In some embodiments, the pharmaceutical product is substantially free of staphylococcus aureus. In certain embodiments, the pharmaceutical product contains less than 10 -1 CFU/mL staphylococcus aureus. In certain embodiments, the pharmaceutical product contains less than 10 - 2 CFU/mL staphylococcus aureus. In certain embodiments, the pharmaceutical product contains less than 10 -3 CFU/mL staphylococcus aureus. In certain embodiments, the pharmaceutical product contains less than 10 -4 CFU/mL staphylococcus aureus. In certain embodiments, the pharmaceutical product contains less than 10 -5 CFU/mL staphylococcus aureus. In certain embodiments, the pharmaceutical product contains less than 10 -6 CFU/mL staphylococcus aureus.
In some embodiments, the pharmaceutical product is substantially free of geobacillus stearothermophilus. In certain embodiments, the pharmaceutical product contains less than 10 -1 CFU/mL Geobacillus stearothermophilus. In certain embodiments, the pharmaceutical product contains less than 10 -2 CFU/mL Geobacillus stearothermophilus. In certain embodiments, the pharmaceutical product contains less than 10 -3 CFU/mL Geobacillus stearothermophilus. In certain embodiments, the pharmaceutical product contains less than 10 -4 CFU/mL Geobacillus stearothermophilus. In certain embodiments, the pharmaceutical product contains less than 10 -5 CFU/mL Geobacillus stearothermophilus. In certain embodiments, the pharmaceutical product contains less than 10 -6 CFU/mL Geobacillus stearothermophilus.
In some embodiments, the pharmaceutical product is substantially free of bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -1 CFU/mL Bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -2 CFU/mL Bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -3 CFU/mL Bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -4 CFU/mL Bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -5 CFU/mL Bacillus pumilus. In certain embodiments, the pharmaceutical product contains less than 10 -6 CFU/mL Bacillus pumilus.
In some embodiments, the pharmaceutical product has at least 10 -3 -10 -6 Sterility assurance level of (c). In certain embodiments, the pharmaceutical product has at least 10 -3 Sterility assurance level of (c). In other embodiments, the pharmaceutical product has at least10 -4 Sterility assurance level of (c). In other embodiments, the pharmaceutical product has at least 10 -5 Sterility assurance level of (c). In other embodiments, the pharmaceutical product has at least 10 -6 Sterility assurance level of (c).
In some embodiments, the pharmaceutical product is resistant to microbial growth. In some embodiments, the pharmaceutical product is chemically stable.
In particular embodiments, the pharmaceutical product as described herein is administered to a subject via direct injection. In other specific embodiments, the pharmaceutical product as described herein is administered to a subject via indirect injection.
In some embodiments, the invention provides a pharmaceutical product as described herein, wherein the pharmaceutical product is packaged in the form of a kit comprising the pharmaceutical product and a syringe.
In some embodiments, the present invention provides pharmaceutical products and packages as described herein, wherein the pharmaceutical products and packages are sterilized by gamma irradiation. In some embodiments, the present invention provides a pharmaceutical product and a package as described herein, wherein the pharmaceutical product is sterilized by ion intensity and gamma irradiation, and the package is sterilized by gamma irradiation. In a specific embodiment, the pharmaceutical product and package are sterilized by gamma irradiation of at least 20 kGy. In some embodiments, the pharmaceutical product and package are sterilized by gamma irradiation of 18-25 kGy. In some embodiments, the pharmaceutical product and package are sterilized by gamma irradiation of 25-33 kGy. In some embodiments, the pharmaceutical product and package are sterilized by gamma irradiation of 45-59 kGy.
In some embodiments, the invention provides a method of treating choline deficiency in a subject comprising administering to the subject an effective amount of a pharmaceutical product as described herein.
In some embodiments, the invention provides methods of providing parenteral support to a subject comprising administering to the subject an effective amount of a pharmaceutical product as described herein.
In some embodiments, the invention provides methods of providing parenteral nutrition to a subject comprising administering to the subject an effective amount of a pharmaceutical product as described herein.
In some embodiments, the invention provides a method of treating liver and gall juice stasis in a subject comprising administering to the subject an effective amount of a pharmaceutical product as described herein.
In some embodiments, the invention provides a method of treating liver steatosis in a subject, comprising administering to the subject an effective amount of a pharmaceutical product as described herein.
In some embodiments, the invention provides a method of treating an intestinal failure-related liver disease (IFALD) in a subject, comprising administering to the subject an effective amount of a pharmaceutical product as described herein.
In some embodiments, the invention provides a method of treating fatty liver disease in a subject comprising administering to the subject an effective amount of a pharmaceutical product as described herein. In some embodiments, the fatty liver disease is AFL, ASH, NAFL, NASH, NASH-associated liver fibrosis or ASH-associated liver fibrosis. In some embodiments, the fatty liver disease is Alcoholic Fatty Liver (AFL). In some embodiments, the fatty liver disease is Alcoholic Steatohepatitis (ASH). In some embodiments, the fatty liver disease is non-alcoholic fatty liver disease (NAFL). In some embodiments, the fatty liver disease is nonalcoholic steatohepatitis (NASH). In some embodiments, the fatty liver disease is NASH-related liver fibrosis. In some embodiments, the fatty liver disease is ASH-related liver fibrosis.
In some embodiments, the invention provides a method of treating choline deficiency in a subject comprising administering to the subject an effective amount of a composition or pharmaceutical product as described herein. In some embodiments, the invention provides a method of treating choline deficiency in a subject comprising administering to the subject an effective amount of a composition as described herein. In some embodiments, the invention provides a method of treating choline deficiency in a subject comprising administering to the subject an effective amount of a pharmaceutical product as described herein.
In some embodiments, the method of treating choline deficiency in a subject further comprises providing parenteral support or parenteral nutrition to the subject. In some embodiments, the method of treating choline deficiency in a subject further comprises providing parenteral support to the subject. In some embodiments, the method of treating choline deficiency in a subject further comprises providing parenteral nutrition to the subject.
In some embodiments, the choline deficiency in the subject is associated with liver and gall juice stasis or liver steatosis. In some embodiments, the choline deficiency in the subject is associated with liver and gall juice stasis. In some embodiments, the choline deficiency in the subject is associated with liver steatosis. In some embodiments, the choline deficiency in the subject is associated with an intestinal failure-related liver disease (IFALD). In some embodiments, the choline deficiency in the subject is associated with a fatty liver disease. In some embodiments, the choline deficiency in the subject is associated with a fatty liver disease, wherein the fatty liver disease is AFL, ASH, NAFL, NASH, NASH-associated liver fibrosis or ASH-associated liver fibrosis. In some embodiments, the choline deficiency in the subject is associated with a fatty liver disease, wherein the fatty liver disease is AFL. In some embodiments, the choline deficiency in the subject is associated with a fatty liver disease, wherein the fatty liver disease is ASH. In some embodiments, the choline deficiency in the subject is associated with a fatty liver disease, wherein the fatty liver disease is NAFL. In some embodiments, the choline deficiency in the subject is associated with a fatty liver disease, wherein the fatty liver disease is NASH. In some embodiments, the choline deficiency in the subject is associated with a fatty liver disease, wherein the fatty liver disease is NASH-associated liver fibrosis. In some embodiments, the choline deficiency in the subject is associated with a fatty liver disease, wherein the fatty liver disease is ASH-associated liver fibrosis.
In some embodiments, the present invention provides a method of synthesizing choline chloride comprising introducing gaseous trimethylamine under pressure with 2-chloroethanol in a hydrogenator in the presence of pure ethanol and methyl tert-butyl ether, wherein the process is a discontinuous process.
In some embodiments, choline chloride is prepared in >99% purity. In some embodiments, choline chloride is prepared in a purity of > 99.5%. In some embodiments, choline chloride is prepared in a purity of > 99.8%.
In some embodiments, the choline chloride produced is substantially free of detectable 2-chloroethanol. In some embodiments, the choline chloride prepared is substantially free of detectable aluminum. In some embodiments, choline chloride is prepared with less than or equal to 0.05ug/g aluminum. In some embodiments, choline chloride is prepared with less than or equal to 0.1% wt. trimethylamine.
Abbreviations (abbreviations)
The following abbreviations are used in the examples, while other abbreviations have the meanings customary in the art:
MTBE: methyl tert-butyl ether
WFI: water for injection
g: gram (g)
L: lifting device
mL: milliliters of (milliliters)
mol: molar (mol)
min: minute (min)
h or hr: hours of
M: mol/L or "molar"
C: degree centigrade
CFU: colony forming units
TNTC: cannot count so much
kGy: thousand gray
USP: united states pharmacopoeia
RH: relative humidity of
VLDL: very low density lipoproteins
PC: phosphatidylcholine
PEMT: phosphatidylethanolamine methyltransferase
DNA: deoxyribonucleic acid
MRI: magnetic resonance imaging
IF: intestinal failure
PN: parenteral nutrition
IFALD: liver disease associated with intestinal failure
IV: intravenous injection
ESLD: advanced liver disease
PNALD: PN related liver disease
NAFL: nonalcoholic fatty liver disease
AFL: alcoholic fatty liver
NASH: nonalcoholic steatohepatitis
ASH: alcoholic steatohepatitis
DC: hierarchically controlled
TSB: pancreas protein soybean broth
TSA: pancreas protein soybean agar
NLT: not less than
NMT: no more than
RRT: relative retention time
RH: relative humidity of
HIAC: high precision
LC (liquid crystal): liquid chromatography
TS: terminal sterilization
API: active pharmaceutical ingredient
FTIR: fourier transform infrared spectroscopy
GC-MS: gas chromatography-mass spectrometry
ICP-MS: inductively coupled plasma mass spectrometry
IC: ion chromatography
QL: quantitative limit
Examples
Embodiments of the invention are further illustrated by the following examples. The following examples are non-limiting and merely represent various aspects of embodiments of the present invention.
Example 1
Preparation of choline salt compositions
Choline chloride (500 g) was combined with WFI and dissolved and made up to 1L in volume, resulting in 50% (w/v) choline chloride in WFI.
Example 2
Sterilization of choline salt compositions by ionic strength
Most single-cell organisms maintain normal ionic strength of about 0.3M within their cells. Any bacteria in a medium with an ionic strength higher than 0.3M will absorb water from the inside of the bacteria until the inside and outside are equal. Bacteria cannot survive normally with little to no water in their cell membranes. Thus, ionic strength can be used as a means to reduce the bacterial population in a medium, thereby sterilizing the medium. Here, 50% (w/v) of the choline chloride in WFI has an ionic strength of about 7M, which may have a bactericidal effect. However, 50% (w/v) of the ionic strength of choline chloride in WFI is insufficient to sterilize organisms such as Geobacillus stearothermophilus, bacillus pumilus, and Staphylococcus aureus (S.aureous). Thus, other sterilization methods, such as heating and gamma irradiation, have been studied.
Example 3
Sterilization of choline salt compositions by heating
D 121 The values were determined for Geobacillus stearothermophilus spores suspended in 50% choline chloride composition. Inoculating 10 each of the vials containing 50% (w/v) choline chloride in WFI 6 The Geobacillus stearothermophilus spores were sealed. Initially (initial population=1.3031x10 6 Individual spores) and after completion of the study (final population = 1.1813x10 6 Spores), microbiological assays were performed on inoculated vials. These assays verify that the spore concentration in the vials is stable for the duration of the study.
Geobacillus stearothermophilus spores D suspended in 50% choline chloride composition 121 The value determinations were determined using spore population data from unexposed vials and vials exposed for 30.0, 60.0, 90.0, and 120.0 minutes. The vials were exposed to Joslyn/Steris Sterilizer corp. Steam b.i.e.r. at 121 ℃. Population assays are then performed to determine spore populations in terms of Colony Forming Units (CFUs). At each exposure time, four repeat units were tested in the same dilution assay. Thus, the plate counts listed below are for each assay (every four units), while the final population is listed as spores for each unit. A spore population of each vial corresponding to each exposure time was used (see table 1,bold numbers used to calculate the final population under the guidance of USP section 55), creates a survival curve, and calculates D according to the survival curve method 121 Values (see figure 1).
TABLE 1
CFU count data and population data for choline chloride injections exposed to vials at 121 °c
The stability of Geobacillus stearothermophilus spores in choline chloride injection suspended in vials was demonstrated. The final population is greater than 1.0x10 6 Individual spores/vials. D of Geobacillus stearothermophilus spores suspended in 50% (w/v) choline chloride in WFI was found 121 The value was 34.0 minutes. D (D) 121 A value of 34 minutes will result in a sterilization cycle of more than 7 hours. A sterilization cycle of 7 hours may lead to degradation or formation of the degradation product. Thus, terminal sterilization with steam is not feasible.
Example 4
Gamma-ray irradiation of choline salt compositions
Due to very high D 121 Values (34 minutes) render sterilization by terminal sterilization impractical, so 50 (w/v) choline chloride injection used in WFI evaluated the feasibility of gamma irradiation to sterilize choline salt compositions. The results of the analysis of the gamma irradiation feasibility study are summarized in table 2.
Briefly, vials and glassware are heat treated to ensure sterility. The initial pH of 50% (w/v) choline chloride in WFI solution was determined. By inoculating Geobacillus stearothermophilus (10) 6 Individual spores) or bacillus pumilus (10) 6 Individual spores) were prepared. The vials were irradiated at gamma doses of 25-33kGy (kilogray) or 45-59 kGy. The control vials were not exposed to gamma irradiation.
TABLE 2
Analysis results of gamma ray irradiation feasibility
1 A pure sample is taken of the sample, 2 1:10 diluted sample
The results in table 2 show that gamma irradiation doses of 45-59kGy produced higher impurities (known and unknown) in the drug product than the control. The gamma irradiation dose of 25-33kGy produced a slightly higher percentage of the known impurity triethylamine in the drug product than the control, but was below the identification limit of the unknown impurity. Based on these results, it was determined that the optimal range for sterilization of 50% (w/v) choline chloride injection in WFI was 25-33kGy gamma dose.
Example 5
Sterilization of choline salt compositions by gamma irradiation
I. Gamma-ray radiation sterilization of geobacillus stearothermophilus/bacillus pumilus
Gamma irradiation was tested as a method of sterilizing 50% (w/v) choline chloride in WFI compositions inoculated with geobacillus stearothermophilus or bacillus pumilus. Vials containing the composition were inoculated with 0.1mL of a suspension of geobacillus stearothermophilus spores, which resulted in 10 per vial 6 A population of CFUs. In this way, individual vials were inoculated with bacillus pumilus. The vials were sterilized using gamma irradiation using 25-33kGy or 45-59 kGy. Vials were selected for use as positive controls without exposure (see table 4). All vials were then assayed to determine spore populations according to CFUs (see table 3). Additional vials were also used for validation of the population assay test (see table 5).
Briefly, sterile vials were tested for sterility using a filtration method. After vortexing, the contents of each vial were removed using a sterile needle and syringe and filtered through its own filter (e.g., 0.2 or 0.45 μm (micrometer) filter). Each vial was used with the same needle, syringe and filter, and the vial was then rinsed, vortexed, and the contents filtered as described above. The process is then repeated to ensure that all residual product is removed from each vial. After filtering the product and vial of rinse, each filter was then rinsed and inoculated onto Tryptic Soy Agar (TSA). Positive controls were tested for each organism type by flushing the filter three times and adding 0.1mL of spore suspension for inoculation to the last aliquot. All plates were incubated according to organism type, wherein Geobacillus stearothermophilus was incubated at 55-60℃and Bacillus pumilus was incubated at 30-35℃for not less than 48 hours (see Table 3).
TABLE 3 Table 3
Exposed test sample-sterility test results
Test organism (Exposure (kGy)) Test sample (CFU)
Bacillus pumilus (25-33 kGy) 0
Bacillus pumilus (25-33 kGy) 0
Bacillus pumilus (25-33 kGy) 0
Bacillus pumilus (45-59 kGy) 0
Bacillus pumilus (45-59 kGy) 0
Bacillus pumilus (45-59 kGy) 0
Geobacillus stearothermophilus (25-33 kGy) 0
Geobacillus stearothermophilus (25-33 kGy) 0
Geobacillus stearothermophilus (25-33 kGy) 0
Geobacillus stearothermophilus (45-59 kGy) 0
Geobacillus stearothermophilus (45-59 kGy) 0
Geobacillus stearothermophilus (45-59 kGy) 0
Negative control (25-33 kGy) 0
Negative control (45-59 kGy) 0
Positive control TNTC
Positive control TNTC
The population and vials for positive controls were tested using a serial dilution protocol using a pour plate methodPurity (see figure 2). Briefly, after vortexing the vials, 1mL of each vial was removed using a sterile needle and syringe (i.e., containing choline chloride solution and 10 5 CFU/mL、10 4 CFU/mL、10 3 CFU/mL or 10 2 CFU/mL vial) and 4 transferred to a test tube containing 9mL of sterile water. The test tube is heat shocked (e.g., 95-100℃for Geobacillus stearothermophilus for 15 minutes and 65-75℃for Bacillus pumilus for 15 minutes). After heat shock, each tube was cooled and vortexed in an ice bath. After cooling, 1mL was transferred to a test tube containing 9mL of sterile water and vortexed (repeated twice). These dilutions resulted in 10 3 CFU/mL、10 2 CFU/mL or 10 1 Concentration of CFU/mL. Will come from 10 2 And 10 1 1mL of the dilution was inoculated into a sterile petri dish, and 18mL of melted TSA was poured into the petri dish and allowed to solidify. This was done for all three (3) positive control vials for each organism type. All plates were incubated according to organism type, with Geobacillus stearothermophilus at 55-60℃and Bacillus pumilus at 30-35℃for not less than 48 hours. After incubation, plates were read, validated, and colonies were reported as CFU/plates (see table 4).
TABLE 4 Table 4
Population assay for unexposed positive control vials
Test organism (Exposure (kGy)) Test sample (CFU)
Bacillus pumilus (10) 1 ) 4
Bacillus pumilus (10) 1 ) 8
Bacillus pumilus (10) 1 ) 7
Bacillus pumilus (10) 2 ) 56
Bacillus pumilus (10) 2 ) 64
Bacillus pumilus (10) 2 ) 75
Geobacillus stearothermophilus (10) 1 ) 2
Geobacillus stearothermophilus (10) 1 ) 2
Geobacillus stearothermophilus (10) 1 ) 4
Geobacillus stearothermophilus (10) 2 ) 28
Geobacillus stearothermophilus (10) 2 ) 25
Geobacillus stearothermophilus (10) 2 ) 37
Negative control 0
Negative control 0
To excite the population assay procedure, a positive control was run. Using a sterile needle and syringe, 0.1mL of the Geobacillus stearothermophilus spore suspension was labeled in one (1) product vial. This inoculation produced 10 per vial 6 A population of CFUs. The above procedure was repeated using a Bacillus pumilus spore suspension. For both organism types, the population assay procedure described above for the unexposed positive control vials was completed. The results are recorded in table 5.
TABLE 5
Verification of population assay test
Test organism (Exposure (kGy)) Test sample (CFU)
Bacillus pumilus (10) 1 ) 169
Bacillus pumilus (10) 1 ) 174
Bacillus pumilus (10) 2 ) TNTC
Bacillus pumilus (10) 2 ) TNTC
Geobacillus stearothermophilus (10) 1 ) 22
Geobacillus stearothermophilus (10) 1 ) 21
Geobacillus stearothermophilus (10) 2 ) 213
Geobacillus stearothermophilus (10) 2 ) 183
Negative control 0
Negative control 0
Vials exposed to gamma irradiation using 25-33kGy or 45-59kGy showed no bacterial growth (cfus=0 for all samples). These data show the effectiveness of gamma irradiation as a 50% (w/v) choline chloride sterilization process in WFI compositions. The unexposed positive control samples showed a lower recovered population than the samples initially inoculated in the vials. The vials tested for validation of the population assay test produced CFUs that were lower than expected, which may indicate that organisms may decrease over time while in solution, with different results/conditions. Population assays performed on the test control showed much higher recovery than the positive control vials, indicating that the method for inoculation is acceptable. All the results show that gamma irradiation is an effective method for sterilizing 50% (w/v) choline chloride in WFI compositions from a microbiological point of view against organisms such as geobacillus stearothermophilus and bacillus pumilus.
Gamma-ray sterilization of staphylococcus aureus
Gamma irradiation was tested as a method of sterilizing 50% (w/v) choline chloride composition in WFI against s. With 0.1mL of Staphylococcus aureus spore suspension (10 9 CFU/mL) was inoculated with a vial (5 mL) containing the composition. The vials were sterilized using gamma radiation of 25-33 kGy. Vials were selected for use as positive controls without exposure. All vials were then assayed to determine spore populations according to CFUs (see table 7). Additional vials were also used for validation of the population assay test (see table 6).
The microorganism retention was measured as follows. Briefly, stock solutions of staphylococcus aureus were prepared in 15mL of Tryptic Soy Broth (TSB) using 10-100CFU organisms and incubated at 30-35 ℃ for about 72 hours, yielding about 10 9 CFU/mL (using McFarland standard approximation No. 4) culture. By combining 10 3 And 10 2 Dilutions were inoculated into TSA in duplicate and incubated at 30-35℃for no more than 24 hours, and population verification was tested in duplicate dilution series of stock cultures prepared. 5mL product vials were labeled with 0.1mL of prepared stock culture. The vials of the labeled product were serially diluted in duplicate using 0.1mL of the labeled product to 9.9mL of sterile water and then 1mL of the diluent was added to 9mL of sterile water. Two dilutions were inoculated in duplicate to TSA using 0.1mL and incubated at 30-35℃for no more than 24 hours. Serial dilutions of the same labeled product vials were tested in 24 hour, 72 hour and 7 day time increments as described above. The results of the microorganism retention can be seen in table 6.
TABLE 6
Results of microorganism retention
Sample of Dilution 1 (10) 3 )CFU Dilution 2 (10) 2 )CFU
Stock cultures (T: 0 hr) TNTC/TNTC 300/100
Product dilution (T: 0 hr) TNTC/TNTC 430/480
Product dilution (T: 24 hr) TNTC/TNTC 430/440
Product dilution (T: 72 hr) TNTC/TNTC 490/550
Product dilution (T: 7 days) TNTC/TNTC 255/266
The sterility results of staphylococcus aureus are shown below. Stock culture of Staphylococcus aureus was performed in 15mL TSB using 10-100CFU organisms and incubated at 30-35℃for about 72 hours, yielding about 10 9 CFU/mL (using McFarland standard approximation No. 4) culture. By combining 10 3 And 10 2 Dilutions were inoculated in duplicate to TSA and incubated at 30-35℃for no more than 24 hours, and population verification was tested in duplicate dilution series of stock cultures prepared. Three will be5mL product vials were labeled with 0.1mL of prepared stock culture. Two additional product vials were labeled with 0.1mL of stock culture to serve as positive controls. Two vials remained uninoculated as negative controls. These vials were sterilized using 25-33kGy gamma irradiation (positive and negative controls remained unexposed). After sterilization, three exposed vials were tested and negative controls were tested using a filtration method. The contents of each vial were individually filtered through a 0.45 μm MCE filter. The vials were rinsed and filtered with the remaining rinse solution in triplicate aliquots of about 100 mL. Filters were inoculated onto TSA and incubated for 5 days at 30-35 ℃. Serial dilutions of the labeled product vials for positive controls were prepared in duplicate using 0.1mL of the labeled product to 9.9mL of sterile water, then 1mL of the dilution was added to 9mL of sterile water and repeated three times. The final 3 dilutions were inoculated into TSA in duplicate using 0.1mL and incubated for 5 days at 30-35 ℃. The sterility results of staphylococcus aureus are shown in table 7.
TABLE 7
Staphylococcus aureus sterility assay results
Test organism (Exposure (kGy)) Test sample (CFU)
Dilution of stock culture 1 st (10≡3) TNTC/TNTC
Dilution of stock culture 2 nd (10≡2) 132/108
Positive control 1-10A 2 0/0
Positive control 2-10A 2 0/0
Positive control 1-10A 3 0/1
Positive control 2-10A 3 0/2
Positive control 1-10≡4 207/238
Positive control 2-10≡4 292/182
Negative control 1 0
Negative control 2 0
Product vial 1 0
Product vial 2 0
Product vial 3 0
Population verification of the two stock culture inoculums for both microbial retention and sterility study assays confirmed the correct reporting population. The inoculated product vials are capable of exhibiting minimal population-reduced microbial retention over an extended period of time. The microbiological retention study showed a log reduction of zero over the course of one week, while the positive control tested from the sterility determination study showed a log reduction of 2-3 over >5 weeks between inoculation and testing. Terminal sterilization with gamma irradiation was demonstrated to be effective as a result of recovery of 0CFU in all three exposed product vials. The results show that gamma irradiation is an effective method for sterilizing 50% (w/v) choline chloride in WFI compositions from a microbiological point of view against organisms such as staphylococcus aureus. The combination of high ion intensity and gamma irradiation has proven to be very effective for sterilization of aqueous choline chloride compositions.
Example 6
Forced degradation of choline chloride
Choline chloride was exposed to the pressure conditions shown in table 8.
TABLE 8
Treatment of forced degradation studies
Each forced degradation solution was prepared by transferring 25mg choline chloride into a 100mL volumetric flask and then exposing to the appropriate conditions. At the end of the treatment, the acid and base treated samples were neutralized with equal amounts of hydrochloric acid or sodium hydroxide. After equilibration to room temperature, each degradation sample was prepared according to the test method. The treated sample formulation was analyzed (single injection) according to this method, as well as the control, except that the run time was prolonged to allow for potential late eluting degradation peaks. The mass balance of each treatment was checked.
The results of the forced degradation study are shown in table 9. The method is capable of separating known (trimethylamine) and unknown degradation products in the presence of acids, bases, oxidation, light and heat to the extent that they can be accurately quantified. Two unknown impurities (RRT 0.75 and 0.78, about 0.3% each) were observed in the sample at the oxidation pressure, and one unknown impurity (RRT 1.42/1.44,0.14%) was observed in the sample at the acid and base pressures. The chromatographic resolution between the active and nearest elution peak (if present at a level of ≡0.05%) is Not Lower Than (NLT) 1.2, and the degradation peaks of ≡0.05% are distinguished from each other and from choline chloride to the extent that they can be quantified (target resolution NLT 1.2). In addition, the chromatography showed acceptable mass balance under all pressure conditions. Choline chloride exhibits good stability under all pressure conditions, wherein no significant degradation is observed when exposed to maximum pressure conditions. Thus, under the conditions described in example 5 for sterilization via gamma irradiation, choline chloride is not expected to degrade significantly.
TABLE 9
Forced degradation of-%lc and%
Example 7
Stability of choline salt compositions
A 50% (w/v) choline chloride solution in WFI was prepared and transferred to a type I glass vial or a layered controlled (DC) vial and stored for 6 months at 25 ℃/60% Relative Humidity (RH) or 40 ℃/75% RH storage conditions to evaluate stability. The data for the 3 month stability results are summarized in tables 10 to 13. The results show that the stability of the drug product is acceptable under both storage conditions, namely 25 ℃/60% rh and 40 ℃/75% rh.
Table 10
Stability data for glass vial I at 25 ℃/60% RH
TABLE 11
Stability data for glass vial I at 40 ℃/75% RH
Table 12
Stability data for DC vials at 25 ℃/60% RH
TABLE 13
Stability data for DC vials at 40 ℃/75% RH
Example 8
pH studies of choline salt compositions
A pH study was performed to evaluate the pH of the choline chloride solution after terminal sterilization with heat. A50% w/v choline chloride solution in WFI was prepared according to example 1. Several vial types were also evaluated, including a Schott type I glass vial, a Schott type I plus glass vial, a Schott layered control (DC) glass vial, and a Kimble type I glass molded vial. Briefly, 50% w/v choline chloride solution (5.5 mL) in WFI was filled in each type of vial. Two filled vials of each type were kept as controls and the remaining vials were terminally sterilized at 121 ℃ for 45 minutes. 10% choline chloride Active Pharmaceutical Ingredient (API) in water is the recommended USP method for determining the pH of choline chloride API. Thus, 50% choline chloride diluted 1:5 in WFI solution was also included in the pH study at each time point. At each time point, the contents from 2 vials of each vial type were combined together separately. 2mL of the combined solution was diluted with 8mL of WFI to prepare a 1:5 diluted sample. The remaining 9mL of solution was subjected to the "as is" (no dilution) sample test. For pre-TS (terminal sterilization) samples, 2 reserved vials from each vial type were kept at room temperature until the terminal sterilized samples were ready for testing. All pH checks were then performed simultaneously. At each time point, pH meters were calibrated with pH standards of 4.0, 7.0, and 10.0, and then samples were tested for pH (see table 14). The change in pH over time is plotted in fig. 3 and 4 for each type of vial. Fig. 3 plots the pH results for the "as received" samples, and fig. 4 plots the pH results for the "1:5 diluted" 50% w/v choline chloride solution in WFI before and after terminal sterilization by heating.
TABLE 14
Results of pH studies
The results showed that the pH of the solution drifts upward during the course of the study. The pH of the solution in the type I plus vial did not drift so much compared to the initial pH reading and was stable to measure the pH of the undiluted "as received" sample. Type I tubular vials, DC and molded vials showed an initial upward pH drift and then flattened at the 4 week time point. The type I plus molded vials had higher 1:5 dilution results than the "as received" pH results and were comparable for type I tubular and DC vials, respectively. The pH results for the 1:5 diluted DC vials were stable compared to other vial types.
Example 9
Methods of treatment using choline salt compositions
The choline salt compositions as described in examples 1 to 7 can be used for treating choline deficiency in a subject associated with liver steatosis and/or cholestasis. The choline salt compositions as described in examples 1 to 7 can also be used to treat choline deficiency in subjects associated with liver disease associated with Intestinal Failure (IFALD), fatty liver disease (e.g., alcoholic Fatty Liver (AFL), alcoholic Steatohepatitis (ASH), non-alcoholic fatty liver (NAFL), non-alcoholic steatohepatitis (NASH), NASH-associated liver fibrosis, or ASH-associated liver fibrosis). The choline salt compositions as described in examples 1 to 7 can be used as components of parenteral support. The choline salt compositions as described in examples 1 to 7 can be used as a component of parenteral nutrition.
Example 10
Synthesis of choline chloride
Scheme 1
Choline chloride is synthesized by combining gaseous trimethylamine with 2-chloroethanol under pressure in a hydrogenator in the presence of ethanol and methyl tert-butyl ether.
Example 11
Analysis of Choline chloride
The choline chloride synthesized in example 10 was analyzed. The results for two different batches of choline chloride can be found in table 15. Both batches were analysed three months after production.
TABLE 15
Analysis of Choline chloride
The choline chloride analysis showed that both batches were prepared in high purity (99.5% and 99.8%, respectively). The amount of residual chlorohydrin was below the Quantitative Limit (QL). In addition, the aluminum content was found to be 0.05ug/g choline chloride or less.
The various embodiments described above may be combined to provide further embodiments. All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent publications mentioned in this specification and/or listed in the application data sheet, including, but not limited to, U.S. provisional patent application No. 63/181,858, filed on 4 months 29 of 2021, are incorporated herein by reference in their entirety. Aspects of the embodiments can be modified, if necessary, to employ concepts of the various patents, applications and publications to provide yet other embodiments.
These and other changes can be made to the embodiments in light of the above detailed description. In general, in the following claims, the terms used should not be construed to limit the claims to the specific embodiments disclosed in the specification and the claims, but should be construed to include all possible embodiments along with the full scope of equivalents to which such claims are entitled. Accordingly, the claims are not limited by the present disclosure.

Claims (30)

1. A sterile composition for intravenous injection comprising choline chloride in an aqueous medium, wherein the choline chloride is present in the composition at a level of 25% to 75% choline chloride by weight/volume%.
2. The composition of claim 1, wherein the composition comprises 50% choline chloride by weight/volume%.
3. The composition of claim 1, wherein the aqueous medium is water for injection.
4. The composition of claim 1, wherein the composition is sterilized by gamma irradiation.
5. The composition of claim 1, wherein the gamma irradiation is at least 20kGy.
6. The composition of claim 1, wherein the gamma irradiation is 25-33kGy.
7. The composition of claim 1, wherein the composition has at least 10 -6 Sterility assurance level of (c).
8. The composition of claim 1, wherein the composition contains less than 10 -1 CFU/mL staphylococcus aureus (S.aureus), geobacillus stearothermophilus (G.stearothermophilus), and/or bacillus pumilus (B.pumilus).
9. The composition of claim 1, wherein the composition is preservative-free.
10. The composition of claim 1, wherein the composition contains a preservative.
11. The composition of claim 1, wherein the composition contains at least one amino acid, at least one vitamin, and/or at least one fatty acid.
12. The composition of claim 1, wherein the composition comprises a pharmaceutically acceptable carrier, diluent, or excipient.
13. The composition of claim 1, wherein the composition has an ionic strength of 0.3-7M.
14. The composition of claim 1, wherein the composition has an ionic strength of about 7M.
15. The composition of claim 1, wherein the composition has a pH of about 4-7.
16. The composition of claim 1, wherein the composition is suitable for administration via indirect injection or via direct injection.
17. A method of preparing a sterile choline chloride composition comprising combining choline chloride with water for injection to produce a 50% (w/v) solution having an ion strength of about 7M and a pH of about 4-7, and exposing the solution to gamma radiation to produce at least 10 -6 Sterility assurance level of (c).
18. The method of claim 17, wherein the gamma irradiation is at least 20kGy.
19. The method of claim 17, wherein the gamma irradiation is 25-33kGy.
20. A sterile aqueous choline chloride pharmaceutical product prepared by the process of:
i) Dissolving choline chloride in water for injection to a final concentration of 40% -60% choline chloride by weight/volume%;
ii) filtering the solution through a 0.2 micron filter;
iii) Transferring the solution to a glass vial;
iv) sealing the vial; and
v) sterilizing the pharmaceutical product using gamma irradiation.
21. The sterile aqueous choline chloride pharmaceutical product of claim 20 wherein the pharmaceutical product contains 50% choline chloride by weight/volume%.
22. The sterile aqueous choline chloride pharmaceutical product of claim 20 wherein the pharmaceutical product is filtered through two 0.2 micron filters in series.
23. The sterile aqueous choline chloride pharmaceutical product of claim 20, wherein the gamma irradiation is at least 20kGy.
24. The sterile aqueous choline chloride pharmaceutical product of claim 20, wherein the gamma irradiation is 25-33kGy.
25. A method of treating choline deficiency in a subject comprising administering to the subject an effective amount of the composition of claim 1.
26. The method of claim 25, further comprising providing parenteral support or parenteral nutrition to the subject.
27. The method of claim 25, wherein the choline deficiency is associated with liver and gall juice accumulation or liver steatosis.
28. The method of claim 25, wherein the choline deficiency is associated with an intestinal failure-related liver disease (IFALD).
29. The method of claim 25, wherein the choline deficiency is associated with fatty liver disease.
30. The method of claim 29, wherein the fatty liver disease is AFL, ASH, NAFL, NASH, NASH-associated liver fibrosis or ASH-associated liver fibrosis.
CN202280030643.3A 2021-04-29 2022-04-28 Sterile aqueous choline salt compositions Pending CN117241782A (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US63/181,858 2021-04-29
US17/246,438 2021-04-30
US17/672,549 2022-02-15
US17/672,549 US12083081B2 (en) 2021-04-29 2022-02-15 Sterile aqueous choline salt compositions
PCT/US2022/026693 WO2022232371A1 (en) 2021-04-29 2022-04-28 Sterile aqueous choline salt compositions

Publications (1)

Publication Number Publication Date
CN117241782A true CN117241782A (en) 2023-12-15

Family

ID=89091643

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202280030643.3A Pending CN117241782A (en) 2021-04-29 2022-04-28 Sterile aqueous choline salt compositions

Country Status (1)

Country Link
CN (1) CN117241782A (en)

Similar Documents

Publication Publication Date Title
EP2567700B1 (en) Vitamin-containing nutrition infusion for administration through peripheral vein
KR101716215B1 (en) Ready to be infused gemcitabine solution
CN102525900A (en) Levetiracetam injection and preparation method thereof
US20220378077A1 (en) Ready-To-Use Parenteral Nutrition Formulation
CN112933045B (en) Co-loaded dihydroartemisinin/chloroquine phosphate double-sensitive nano preparation and preparation method thereof
Zhou et al. Using tea nanoclusters as β-lactamase inhibitors to cure multidrug-resistant bacterial pneumonia: a promising therapeutic strategy by Chinese materioherbology
CN103222978A (en) Compound sulphamethoxazole injection and preparation method thereof
CN117241782A (en) Sterile aqueous choline salt compositions
US12083081B2 (en) Sterile aqueous choline salt compositions
BRPI0713639A2 (en) Method for the manufacture of an intravenous or intramuscular artesunic acid formulation, artesunic acid formulation, kit for producing an artesunic acid formulation, methods for treating a patient with malaria, and for sterilization of artesunic acid powder, and of artesunic acid
US11311503B1 (en) Sterile aqueous choline salt compositions
CN102908312B (en) Liquid combination for resisting hepatitis B viruses
CN107920994A (en) The composition and method of lamellar body for therapeutic purposes
CN116056726A (en) Parenteral nutrition containing trace elements
EP3689362A1 (en) Stable compositions of neuroactive peptides
CN103432067A (en) Ketoprofen solution and preparation method thereof
CN104244925B (en) The stability of the raising of dihydromorphinone hydrochloride solution
CN102525910A (en) Process for preparing penehyclidine hydrochloride injection
CN110179810B (en) A pharmaceutical composition with anti-tumor effect
Pandit et al. Exploring the protective effect of Ascorbic acid on Amoxicillin and Clavulanic acid-induced lipid peroxidation in goat liver homogenate
CN110314132B (en) Ornithine aspartate injection and preparation method thereof
Stanley et al. Comparison for pharmaceutical equivalence of FDA-approved products and compounded preparations of ketoprofen, amikacin, and boldenone.
WO2023191082A1 (en) Formulation having improved absorbability of low-soluble and/or low-membrane-permeable drug
RU2395278C1 (en) Method of obtaining complex medication for prevention and treatment of metabolism pathology and malfunctions of immune system in animals
CN101152171B (en) Method of preparing mildronate injection

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
REG Reference to a national code

Ref country code: HK

Ref legal event code: DE

Ref document number: 40105280

Country of ref document: HK